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US 12,466,875 B2Granted
Supramolecular high affinity protein-binding system for purification of biomacromolecules
AssigneesThe Johns Hopkins University, Bristol-Myers Squibb Company
InventorsHonggang Cui, Yi Li, Xuankuo Xu, Lye Lin Lock, Zhengjian Li
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Abstract
In certain embodiments, the present invention provides novel antibody purification methods and systems using a potentially simple and cost-efficient means. In some embodiments, customized Z-33 derived from Staphylococcus aureus Protein A is used to construct immuno-amphiphile molecules which can assemble into immunofibers in aqueous solution with bioactive epitopes on the surface and have IgG binding ability.
Claims11
A method for purifying an antibody or an Fc fusion protein, the method comprising the steps of:
- a) dissolving an immuno-amphiphile in an aqueous solution at a physiological pH to provide a dissolved immuno-amphiphile;
- and aging the dissolved immuno-amphiphile overnight to provide immunofibers (IFs);
- b) mixing a sample containing an antibody or an Fc fusion protein with the IFs, and allowing the IFs to bind the Fc portion of the antibody or Fc fusion protein to form an immunofiber-antibody complex or immunofiber-Fc fusion protein complex in a solution;
- c) separating the immunofiber-antibody complex or the immunofiber-Fc fusion protein complex from the solution by adding salt and centrifugation;
- and
- d) dissociating the IFs from the antibody or Fc fusion protein and collecting the unbound antibody or Fc fusion protein,
- wherein the immuno-amphiphile comprises an antibody binding peptide and a hydrocarbon moiety;
- wherein the hydrocarbon moiety is conjugated to the N-terminus of the antibody binding peptide;
- wherein the antibody binding peptide consists of the amino acid sequence FNMQQQRRFYEALHDPNLNEEQRNAKIKSIRDD (SEQ ID NO:
- 1);
- and
- wherein the hydrocarbon moiety is a C12 hydrocarbon chain or two C8 hydrocarbon chains.
9 more elements
Dependencies are inferred from the claim text.